Polyvinyl alcohol/poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PVA/PEDOT:PSS) composite ultrafine fibers were successfully fabricated by high pressure airflow assisted electrospinning. The electrical properties of PVA/PEDOT:PSS nanofibers with different diameters were characterized. The average diameter of the nanofibers can be down to 68 nm. Due to its large specific surface area, ammonia sensing of the ultrafine nanofibers is more sensitive than the traditional electrospun fibers (average fiber diameter of 263 nm). The ammonia sensing properties of the samples were tested by impedance analysis. The results show that ultrafine PVA/PEDOT:PSS nanofibers are more suitable for detecting low concentrations of ammonia with higher sensitivity.